The clinical implications of recent studies on the structure and function of the retinal microvasculature in diabetes.
The retinal blood vessels provide the opportunity to study early structural and functional changes in the microvasculature prior to clinically significant microvascular and macrovascular complications of diabetes. Advances in digital retinal photography and computerised assessment of the retinal vasculature have provided more objective and precise measurements of retinal vascular changes. Clinic- and population-based studies have reported that these quantitatively measured retinal vascular changes (e.g. retinal arteriolar narrowing and venular widening) are associated with preclinical structural changes in other microvascular systems (e.g. infarct in the cerebral microcirculation), as well as diabetes and diabetic complications, suggesting that they are markers of early microvascular dysfunction. In addition, there are new retinal imaging techniques to further assess alterations in retinal vascular function (e.g. flicker-induced vasodilatory response, blood flow and oxygen saturation) in diabetes and complications that result from the effects of chronic hyperglycaemia, inflammation and endothelial dysfunction. In this review, we summarise the latest findings on the relationships between quantitatively measured structural and functional retinal vascular changes with diabetes and diabetic complications. We also discuss clinical implications and future research to evaluate whether detection of retinal vascular changes has additional value beyond that achieved with methods currently used to stratify the risk of diabetes and its complications.


Bond strength of luting materials to ceramic crowns after different surface treatments.
The aim of present study was to evaluate the effect of different pre-treatments of alumina and zirconia copings on the bond strength of different luting materials. Pull out tests was performed on 60 alumina and 80 zirconia copings. Randomly selected, copings were divided in groups of i) un-treated alumina and zirconia copings, (n=20) ii) alumina and zirconia copings sandblasted with 50 or 110 microm alumina particles respectively (n=20), iii) zirconia copings treated with monolayer of glass pearls fused to the inner surface (n=20), iv) zirconia copings treated with silanized glass pearls (n=10). Zinc phosphate, Panavia 21 and VarioLink II were used for cementation. Pull out tests were done in an Instron universal testing machine with a speed of 0.5 mm/min and fracture loads was measured in N. Untreated zirconia copings cemented with zinc phosphate showed significantly higher bond strength values compared to those with sandblasted surfaces. No difference was seen between untreated alumina copings and those with sandblasted surfaces. Sandblasting decreased bond strength of zirconia and alumina copings. Glass pearls increased bond strength of zirconia copings, which was even better after silanization. Variolink II in combination with alumina gave significantly lower bond strength.


Computer programs as theories in biology.
Complex theories in biology may be developed, refined, and tested by the use of computer programmed simulations. The computer is recognized as a powerful tool for theory development; it is, in fact, the only means of thoroughly testing and examining a large and intricate theory. A program as a text is a statement of a theory and when run on the computer it is model of that theory. As the program's behavior is then the major argument for the credibility of a large and complex theory, the program itself is the only irrefutable statement of the theory. Bur programs written in the currently available programming languages tend to be incomprehensible. We argue that the program should be the definitive statement of the theory. In addition, the program plus a series of abstractions is a vehicle for effective communication of complex theories in biology. Several techniques of computer science are borrowed, for the purpose of developing a methodology for abstraction and a language for representing abstractions. The arguments are fully illustrated with a recently published biological theory.


Gamma interferon production and plasma concentrations of pregnancy-associated glycoproteins 1 and 2 in gestating dairy cows naturally infected with Neospora caninum.
Gamma interferon (IFN-γ) production and cross-breed pregnancy have been attributed a role in protecting dairy cows infected with Neospora caninum against abortion. Plasma levels of pregnancy-associated glycoproteins-1 (PAG-1) are a marker of placental/foetal well-being and of PAG-2 is an abortion risk indicator in chronically N. caninum-infected animals. The present study examines, in cross-breed pregnancies, interactions between IFN-γ production and levels of PAG-1 and PAG-2 in non-aborting naturally Neospora-infected dairy cows. Data were obtained from 60 pregnant Holstein-Friesian cows: 44 Neospora-seropositive and 16 Neospora-seronegative; 12 became pregnant using Holstein-Friesian semen and 48 using Limousin semen. Blood samples were collected on Days 40, 90, 120, 150, 180 and 210 of gestation. Gamma interferon was only detected in the plasma of nine of the 44 Neospora-seropositive cows, all of them became pregnant using Limousin semen. Through GLM procedures, in cows inseminated with Limousin semen and Neospora-seropositive cows showing no IFN-γ production, PAG-1 concentrations were high and increased throughout gestation compared to the levels detected in cows inseminated with Holstein-Friesian semen and Neospora-seropositive cows producing IFN-γ, respectively. In Neospora-seronegative cows and in Neospora-seropositive cows showing no IFN-γ production, significantly increased PAG-2 concentrations were observed on gestation Day 120. Our findings indicate that IFN-γ production correlates negatively and the production of antibodies against N. caninum is uncorrelated with plasma PAG concentrations during gestation in Neospora-infected dairy cows. Accordingly, IFN-γ production could be linked to the transplacental migration of tachyzoites, which may cause a reduction in PAG levels.


Protease-targeting peptide-functionalized porous silicon nanoparticles for cancer fluorescence imaging.
Background: Porous silicon (pSi) nanoparticles (NPs) functionalized with suitable targeting ligands are now established cancer bioimaging agents and drug-delivery platforms. With growing interest in peptides as tumor-targeting ligands, much work has focused on the use of various peptides in combination with pSi NPs for cancer theranostics. Here, the authors investigated the targeting potential of pSi NPs functionalized with two types of peptide, a linear 10-mer peptide and its branched (Y-shaped) equivalent, that respond to legumain activity in tumor cells. Results: In vitro experiments established that the linear peptide-pSi NP conjugate had better aqueous stability under tumor conditions and higher binding efficiency (p < 0.001) toward legumain-expressing cells such as RAW 264.7 cells compared with that of its branched equivalent. In vivo studies (analyzed using ex vivo fluorescence) with the linear peptide-pSi NP formulation using a syngeneic mouse model of breast cancer showed a higher accumulation (p > 0.05) of linear peptide-conjugated pSi NPs in the tumor site within 4 h compared with nonconjugated pSi NPs. These results suggest that the linear peptide-pSi NP formulation is a nontoxic, stable and efficient fluorescence bioimaging agent and potential drug-delivery platform.


Emergent Elasticity in Amorphous Solids.
The mechanical response of naturally abundant amorphous solids such as gels, jammed grains, and biological tissues are not described by the conventional paradigm of broken symmetry that defines crystalline elasticity. In contrast, the response of such athermal solids are governed by local conditions of mechanical equilibrium, i.e., force and torque balance of its constituents. Here we show that these constraints have the mathematical structure of a generalized electromagnetism, where the electrostatic limit successfully captures the anisotropic elasticity of amorphous solids. The emergence of elasticity from local mechanical constraints offers a new paradigm for systems with no broken symmetry, analogous to emergent gauge theories of quantum spin liquids. Specifically, our U(1) rank-2 symmetric tensor gauge theory of elasticity translates to the electromagnetism of fractonic phases of matter with the stress mapped to electric displacement and forces to vector charges. We corroborate our theoretical results with numerical simulations of soft frictionless disks in both two and three dimensions, and experiments on frictional disks in two dimensions. We also present experimental evidence indicating that force chains in granular media are subdimensional excitations of amorphous elasticity similar to fractons.


Investigating a rare methicillin-resistant Staphylococcus aureus strain: first description of genome sequencing and molecular characterization of CC15-MRSA.
Methicillin resistant Staphylococcus aureus CC15 strains (CC15-MRSA) have only been sporadically described in literature. This study was carried out to describe the genetic make-up for this rare MRSA strain. Four CC15-MRSA isolates collected in Riyadh, Saudi Arabia, between 2013 and 2014 were studied. Two isolates were from clinical infection and 2 from retail meat products. Whole genome sequencing was carried out using Illumina HiSeq2500 genome analyzer. All the CC15-MRSA isolates had the multilocus sequence typing profile ST1535, 13-13-1-1-81-11-13, which is a single locus variant of ST15. Of the 6 contigs related to the SCC element, one comprised a recombinase gene ccrAA, ccrC-PM1, fusC and a helicase, another one included mvaS, dru, mecA and 1 had yobV and Q4LAG7. The SCC element had 5 transposase genes, namely 3 identical paralogs of tnpIS431 and 2 identical paralogs of tnpIS256. Two identical copies of a tnpIS256-based insertion element flank the aacA-aphD gene. Two copies of this insertion element were present with 1 located in the SCC element and another inserted into the sasC gene. A short 3 kb region, which lacks any bacteriophage structural genes and site-specific DNA integrase, was inserted into the hlb gene. The hsdM and the 5'-part of the hsdS gene are replaced by a copy of the hsdM/hsdS paralogs from νSaβ giving rise to a new chimeric paralog of hsdS in νSaα. CC15-MRSA shows a novel SCCmecV/SCCfus composite element. Its variant of hsdM/hsdS probably facilitated uptake of foreign mobile genetic elements that promoted emergence of CC15-MRSA. Close surveillance is needed to monitor spread and emergence of further CC15 MRSA strains.


Maternal care, hippocampal glucocorticoid receptors, and hypothalamic-pituitary-adrenal responses to stress.
Variations in maternal care affect the development of individual differences in neuroendocrine responses to stress in rats. As adults, the offspring of mothers that exhibited more licking and grooming of pups during the first 10 days of life showed reduced plasma adrenocorticotropic hormone and corticosterone responses to acute stress, increased hippocampal glucocorticoid receptor messenger RNA expression, enhanced glucocorticoid feedback sensitivity, and decreased levels of hypothalamic corticotropin-releasing hormone messenger RNA. Each measure was significantly correlated with the frequency of maternal licking and grooming (all r's > -0.6). These findings suggest that maternal behavior serves to "program" hypothalamic-pituitary-adrenal responses to stress in the offspring.


An unusual cluster of circumscribed juvenile pityriasis rubra pilaris cases.
Circumscribed juvenile pityriasis rubra pilaris (PRP) is an uncommon dermatosis. We describe the unusual clustering of circumscribed juvenile PRP cases in our pediatric dermatology clinic in 2011. A retrospective chart review was done of patients presenting during the summer of 2011 with classic findings of circumscribed juvenile PRP. Clinical data including past medical and family history, presenting symptoms, infectious disease history and evaluation, biopsy results, and management were recorded. Seven patients, ages 5 to 19 years, all had strikingly similar skin findings of pink to hyperpigmented, well-defined, scaly papules and plaques on their elbows, knees, dorsal hands, ankles, and Achilles tendons. Four of the seven also had palmoplantar involvement. Four were sibling pairs and the other three were unrelated. Streptococcus pyogenes infection was suspected as a trigger in four of the patients. The unusual clustering of this uncommon disease, along with the occurrence in two sibling pairs, suggests that a genetic susceptibility unmasked by an infectious agent may play a role in its pathogenesis.


Partial amino acid sequences around sulfhydryl groups of soybean beta-amylase.
Sulfhydryl (SH) groups of soybean beta-amylase were modified with 5-(iodoaceto-amidoethyl)aminonaphthalene-1-sulfonate (IAEDANS) and the SH-containing peptides exhibiting fluorescence were purified after chymotryptic digestion of the modified enzyme. The sequence analysis of the peptides derived from the modification of all SH groups in the denatured enzyme revealed the existence of six SH groups, in contrast to five reported previously. One of them was found to have extremely low reactivity toward SH-reagents without reduction. In the native state, IAEDANS reacted with 2 mol of SH groups per mol of the enzyme (SH1 and SH2) accompanied with inactivation of the enzyme owing to the modification of SH2 located near the active site of this enzyme. The selective modification of SH2 with IAEDANS was attained after the blocking of SH1 with 5,5'-dithiobis-(2-nitrobenzoic acid). The amino acid sequences of the peptides containing SH1 and SH2 were determined to be Cys-Ala-Asn-Pro-Gln and His-Gln-Cys-Gly-Gly-Asn-Val-Gly-Asp-Ile-Val-Asn-Ile-Pro-Ile-Pro-Gln-Trp, respectively.


Cerebellar mutism caused by arteriovenous malformation of the vermis.
Transient mutism following posterior fossa tumour resection in children is well known in the literature. To our Knowledge, this phenomenon has never been reported without surgical intervention. We report a case of cerebellar mutism secondary haemorrhage from a vermian arteriovenous malformation (AVM), which resolved to ataxic dysarthria after 6 weeks. Embolization of the AVM was performed and the patient's clinical status continued to improve gradually till she became normal 6 months from the insult. The mutism was due to rupture of the AVM which might correlate the cerebellar mutism with the transient vasospasm of the blood vessels supplying the cerebellum. It is possible for a spontaneus posterior fossa bleed to result in transient mutism similar to post surgical mutism.


Phase I dose-escalation study of brostallicin, a minor groove binder, in combination with cisplatin in patients with advanced solid tumors.
Brostallicin is a DNA minor groove binder which shows enhanced antitumor activity in cells which are resistant to several anticancer agents due to their high glutathione S-transferase (GST)/glutathione content. Phase I and II clinical trials of single-agent brostallicin have shown that myelotoxicity is the dose-limiting toxicity (DLT), while hints of antitumor activity were mainly observed in soft tissue sarcoma. Preclinical studies showing a more than additive antitumor effect of the cisplatin-brostallicin combination paved the way to clinical combination studies. In particular, we set up the first clinical combination study of brostallicin and cisplatin in patients with advanced solid tumors. This study was to be followed by a phase II study in patients with recurrent squamous cell carcinoma of the head and neck (SCCHN). Escalating doses of brostallicin were administered in combination with a fixed dose of cisplatin (75 mg/m(2)) in patients with recurrent or metastatic advanced solid tumors who had previously received a cumulative dose of cisplatin not higher than 475 mg/m(2). The recommended dose of brostallicin was expanded in order to have a better estimate of antitumor activity and to better define the safety profile of the combination. Twenty-one patients were treated. Two DLTs (grade 3 fatigue and febrile neutropenia) were observed at dose level 3 (brostallicin 9 mg/m(2)). Dose level 2 (brostallicin 7 mg/m(2) and cisplatin 75 mg/m(2)) was recommended for future phase II studies. Main toxicity was hematologic; in fact, only 1 patient out of 21 did not develop neutropenia and only 2 patients did not have thrombocytopenia. Grade 3-4 neutropenia was observed in 90.5% of patients, grade 3-4 thrombocytopenia in 38.1%, grade 3-4 anemia in 23.8%. The cycle 1 nadir (ANC < 500 x 10(9)/L) for neutrophils was Day 14 (median; range 11-17) with recovery to an ANC of >1,500 3.5 days after nadir (median; range 2-4) at dose level 3. The cycle 1 nadir (median of 51,000 x 10(9)/L) for platelets occurred on Day 13 (median; range 10-15) with recovery to a platelet count of >100,000 4 days after nadir (median; range 2-8). No objective responses were observed, but seven patients had a long lasting (>18 weeks) stable disease. Further studies of the combination of brostallicin and cisplatin are warranted.


Next-Generation Tools to Study Autonomic Regulation In Vivo.
The recent development of tools to decipher the intricacies of neural networks has improved our understanding of brain function. Optogenetics allows one to assess the direct outcome of activating a genetically-distinct population of neurons. Neurons are tagged with light-sensitive channels followed by photo-activation with an appropriate wavelength of light to functionally activate or silence them, resulting in quantifiable changes in the periphery. Capturing and manipulating activated neuron ensembles, is a recently-designed technique to permanently label activated neurons responsible for a physiological function and manipulate them. On the other hand, neurons can be transfected with genetically-encoded Ca2+ indicators to capture the interplay between them that modulates autonomic end-points or somatic behavior. These techniques work with millisecond temporal precision. In addition, neurons can be manipulated chronically to simulate physiological aberrations by transfecting designer G-protein-coupled receptors exclusively activated by designer drugs. In this review, we elaborate on the fundamental concepts and applications of these techniques in research.


The temperature--dependent expression of GST of Schistosoma japonicum (Philippine strain).
Obtained from pSj5, the cDNA gene encoding GST antigen of Schistosoma japonicum (Philippine strain) was ligated with efficient temperature-dependent PBV220 vector which was constructed in CAPM, and then introduced into host bacterium-DH5 alpha (E. coli) by transformation. Transformants were selected by ampicillin and recombinant clones were identified by restriction mapping. The result showed that recombinant clone 43 was the one carrying recombinant plasmid PBV 220 with the correct insertion of the gene fragment. The GST expression ability of clone 43 was investigated by GST enzymic activity assay and SDS-PAGE. A relatively high level of GST enzymic activity was expressed by this clone under the temperature-dependent condition, that is, cultured at 30 degrees C and expressed at 42 degrees C. A more strongly stained 26 kDa protein band was identified by SDS-PAGE. The result indicated that GST of S. japonicum (Philippine strain) could be expressed not only by IPTG induction, but also by the temperature-dependent method.


Hepatitis B and C seropositivity among nursing students at a Turkish university.
Viral hepatitis infections are major health problems, which are especially prevalent in developing countries. In particular, health care workers are at high risk for contracting viral hepatitis. Therefore, preventive measures to minimize occupational exposure to blood borne pathogens are needed to protect both health care workers and nursing students. To determine the prevalence of Hepatitis B and C infections and risk factors among nursing students. Venous bloods were obtained from nursing students and were serologically tested using the ELISA method. A questionnaire form was also used to obtain data related to demographic and socio-economic characteristics of the participants. 0.7% of the nursing students were serologically positive for HBsAg, 17.0% for Anti-HBs and 7.5% were positive for Anti-HBc (IgG). No statistical relationship was determined between all independent variables and the results of serologic tests.


Low-cost and portable MRI.
Research in MRI technology has traditionally expanded diagnostic benefit by developing acquisition techniques and instrumentation to enable MRI scanners to "see more." This typically focuses on improving MRI's sensitivity and spatiotemporal resolution, or expanding its range of biological contrasts and targets. In complement to the clear benefits achieved in this direction, extending the reach of MRI by reducing its cost, siting, and operational burdens also directly benefits healthcare by increasing the number of patients with access to MRI examinations and tilting its cost-benefit equation to allow more frequent and varied use. The introduction of low-cost, and/or truly portable scanners, could also enable new point-of-care and monitoring applications not feasible for today's scanners in centralized settings. While cost and accessibility have always been considered, we have seen tremendous advances in the speed and spatial-temporal capabilities of general-purpose MRI scanners and quantum leaps in patient comfort (such as magnet length and bore diameter), but only modest success in the reduction of cost and siting constraints. The introduction of specialty scanners (eg, extremity, brain-only, or breast-only scanners) have not been commercially successful enough to tilt the balance away from the prevailing model: a general-purpose scanner in a centralized healthcare location. Portable MRI scanners equivalent to their counterparts in ultrasound or even computed tomography have not emerged and MR monitoring devices exist only in research laboratories. Nonetheless, recent advances in hardware and computational technology as well as burgeoning markets for MRI in the developing world has created a resurgence of interest in the topic of low-cost and accessible MRI. This review examines the technical forces and trade-offs that might facilitate a large step forward in the push to "jail-break" MRI from its centralized location in healthcare and allow it to reach larger patient populations and achieve new uses. Level of Evidence: 5 Technical Efficacy Stage: 6 J. Magn. Reson. Imaging 2019. J. Magn. Reson. Imaging 2020;52:686-696.


Tissue Factor Residues That Modulate Magnesium-Dependent Rate Enhancements of the Tissue Factor/Factor VIIa Complex.
The blood coagulation cascade is initiated when the cell-surface complex of factor VIIa (FVIIa, a trypsin-like serine protease) and tissue factor (TF, an integral membrane protein) proteolytically activates factor X (FX). Both FVIIa and FX bind to membranes via their γ-carboxyglutamate-rich domains (GLA domains). GLA domains contain seven to nine bound Ca(2+) ions that are critical for their folding and function, and most biochemical studies of blood clotting have employed supraphysiologic Ca(2+) concentrations to ensure saturation of these domains with bound Ca(2+). Recently, it has become clear that, at plasma concentrations of metal ions, Mg(2+) actually occupies two or three of the divalent metal ion-binding sites in GLA domains, and that these bound Mg(2+) ions are required for full function of these clotting proteins. In this study, we investigated how Mg(2+) influences FVIIa enzymatic activity. We found that the presence of TF was required for Mg(2+) to enhance the rate of FX activation by FVIIa, and we used alanine-scanning mutagenesis to identify TF residues important for mediating this response to Mg(2+). Several TF mutations, including those at residues G164, K166, and Y185, blunted the ability of Mg(2+) to enhance the activity of the TF/FVIIa complex. Our results suggest that these TF residues interact with the GLA domain of FX in a Mg(2+)-dependent manner (although effects of Mg(2+) on the FVIIa GLA domain cannot be ruled out). Notably, these TF residues are located within or immediately adjacent to the putative substrate-binding exosite of TF.


Functional coupling of rat group II metabotropic glutamate receptors to an omega-conotoxin GVIA-sensitive calcium channel in human embryonic kidney 293 cells.
Metabotropic glutamate receptors are G protein-coupled receptors that perform a variety of modulatory roles in the central and peripheral nervous systems. The development of receptor subtype-specific agonists/antagonists has lagged far behind the isolation and characterization of receptor cDNAs. Further more, the coupling of specific metabotropic receptors to the various neuronal-specific effector molecules, such as voltage gated Ca2+ channels, has not been well studied. It was recently demonstrated that a rat group II metabotropic receptor (rm-GluR2) is capable of coupling to endogenous N-type Ca2+ channels when heterologously expressed in adult rat sympathetic ganglia neurons. To eventually understand the molecular aspects of metabotropic receptor modulation of the N-type Ca2+ channel, we have transiently expressed both group II receptors in a human embryonic kidney 293 cell line (G1A1) that stably expresses the human alpha 1B-1, alpha 2b, and beta 1-3 Ca2+ channel subunits. rmGluR2 and rmGluR3 modulate the omega-conotoxin GVIA-sensitive Ba2+ currents in G1A1 cells using a voltage-dependent mechanism via an endogenous pertussis toxin-sensitive G protein. Cell-attached "macropatch" recordings demonstrate that modulation by rmGluR2 and rmGluR3 is membrane delimited. This is the first report of Ca2+ channel modulation mediated by rmGluR3. In addition, an extensive pharmacological comparison between rmGluR2 and rmGluR3 reveals that these group II receptors interact with agonists and antagonists in unique ways.


Syndromic progressive neurodegenerative disease of infancy caused by novel variants in HIBCH: Report of two cases in Colombia.
3-Hydroxyisobutyryl-coenzyme A (CoA) hydrolase deficiency (HIBCHD; MIM: #250620) is a rare autosomal recessive inborn error of metabolism caused by a defect in the HIBCH enzyme, resulting in a deficiency of the conversion of 3-hydroxy-isobutyryl-CoA to 3-hydroxy-isobutyric acid, a critical step in valine catabolism. This neurodegenerative disease of infancy is associated with hypotonia, developmental delay, cerebral atrophy and lesions in the basal ganglia on magnetic resonance imaging (MRI). In this study, we describe two unrelated patients with infantile-onset progressive neurodegenerative disease and mutations in HIBCH identified using whole exome sequencing (WES). In Case 1, WES revealed a novel homozygous variant in the HIBCH gene: c.808A>G (p.Ser270Gly). In Case 2, a novel compound heterozygous mutation in the HIBCH gene is described: c.808A>G (p.Ser270Gly) and c.173A>G (p. Asn58Ser). Parent analysis revealed that c.808A>G (p.Ser270Gly) was inherited from the father and c.173A>G (p. Asn58Ser) from the mother. These novel mutations were predicted as a disease-causing mutation. Plasma acylcarnitine analysis was normal in both patients. Physical examination showed similar features, such as axial hypotonia and spastic hypertonia in the legs. The first patient presented with difficult-to-treat seizures, while the second patient has not yet experienced documented seizures. In conclusion, our findings would widen the mutation spectrum of HIBCH deficiency and the phenotypic spectrum of the disease. The potential genotype-phenotype correlation would be profitable for the correct diagnosis, treatment and integral management of patients with HIBCH deficiency.


Propylthiouracil-coated biodegradable polymer inhibited neointimal formation and enhanced re-endothelialization after vascular injury.
The drug-eluting stent is a standard approach for the treatment of coronary artery disease. Propylthiouracil (PTU), an antithyroid drug, has been proven to suppress neointimal formation after balloon injury. This study used a biodegradable polymer coating with PTU to test its effects on platelet function, re-endothelialization, and neointimal formation after vascular injury. Electrospinning was used to fabricate hybrid stents and generate PTU-loaded nanofibers. PTU-eluting stents maintained a stable release of PTU for 3 weeks. The PTU-coated stent markedly decreased the neointimal formation induced by vascular injury in the descending aorta of rabbits. Moreover, the PTU coating reduced platelet adhesion on the surface of the biodegradable membrane, which was reflected by the decreased expression of adhesion molecule in PTU-treated endothelial cells. The PTU coating enhanced re-endothelialization in injured aortas. In vitro, PTU exerted less suppressive effect on the proliferation and migration of endothelial cells than on those of vascular smooth muscle cells. Furthermore, treatment of endothelial cells with PTU induced phosphorylation (Ser1177) of endothelial nitric oxide synthase as well as its association with heat shock protein 90, supporting the protective role of PTU in endothelial function. The level of thyroid-stimulating hormone remained unchanged during the experimental period. This study indicates that PTU can be released locally and steadily in injured aortas, with some local effects but without systemic effects. Furthermore, PTU-coated stents may have beneficial effects on neointimal formation, endothelial cell, and platelet functions.


Radiopharmaceuticals for bone malignancy therapy.
This continuing education article reviews radionuclide bone therapy agents that are available commercially and introduces agents that are being evaluated for future use. Currently these agents are used to provide pain palliation from metastases to bone. Future applications may include adjuvant therapy to surgery or external beam treatment. After reading this paper, the reader should be able to: (a) describe the desirable characteristics of radionuclide bone therapy agents; (b) compare and contrast radiopharmaceuticals available for bone therapy; and (c) state the clinical applications of radionuclide bone therapy agents.


Enhancing psychosocial outcomes for young adult childhood CNS cancer survivors: importance of addressing vocational identity and community integration.
The purpose of this study was to examine the relationship between vocational identity, community integration, positive and negative affect, and satisfaction with life in a group of young adult central nervous system (CNS) cancer survivors. Participants in this study included 45 young adult CNS cancer survivors who ranged in age from 18 to 30 years (mean=22, SD=3.62), with a mean age at diagnosis of 8.8 years. Four standardized instruments were used to assess the individual's affect, satisfaction with life, vocational identity, and self-perceived level of community integration. Pearson correlation analyses were carried out to examine the relationships between the satisfaction with life and positive and negative affect, vocational identify, and community integration. A hierarchical linear regression was then performed to determine how well these variables predict satisfaction with life among CNS cancer survivors. Significant positive correlations were found between life satisfaction and positive affect (r=0.423, P<0.01), as well as life satisfaction and community integration (r=-505, P<0.001). A positive correlation between life satisfaction and vocational identity was only marginally significant (r=0.312, P<0.05). Regression results indicate that the model explained 29% of the variance with community integration making a unique contribution. The largest contribution of this study is that the findings provide initial evidence that addressing vocational identity and community integration may be important constructs in improving young adult CNS survivors' overall levels of satisfaction with life. These constructs have not been typically addressed in psychosocial cancer treatment programs.


Effects of washing on the bacterial flora of the stallion's penis.
Six stallions were subjected to extensive cleansing of the penis and prepuce with water, Ivory Soap and water, or Betadine surgical scrub and water. The stallions were all washed for 14 days, and then allowed 14 days respite. This pattern of washing and resting was repeated consecutively. Swabs were taken from all 7 stallions twice weekly and semen was collected once a week for bacteriological examination. All forms of cleansing altered the bacterial flora of the stallion's penis; the Ivory Soap tended to encourage the replacement of the normal flora with coliform organisms, while Betadine favoured the growth of Pseudomonas aeruginosa and Klebsiella spp. This experiment showed that the systematic washing of a stallion's penis will cause the normal flora to be replaced with pathogens and potential pathogens.


Open repair of hiatus hernia: abdominal approach.
Open hiatus hernia repair can be accomplished with very low mortality and excellent long-term results. The vast majority of hiatus hernias, including those with peptic structure, can be repaired transabdominally. The Hill repair, the Nissen, and the Collis-Nissen are well-established techniques for repairing a hiatus hernia that have stood the test of time and are associated with good long-term results. The technical aspects of these repairs are discussed in detail within this article.


Interhemispheric hygroma after decompressive craniectomy: does it predict posttraumatic hydrocephalus?
The aim of this study was to determine the incidence of posttraumatic hydrocephalus in severely head-injured patients who required decompressive craniectomy (DC). Additional objectives were to determine the relationship between hydrocephalus and several clinical and radiological features, with special attention to subdural hygromas as a sign of distortion of the CSF circulation. The authors conducted a retrospective study of 73 patients with severe head injury who required DC. The patients were admitted to the authors' department between January 2000 and January 2006. Posttraumatic hydrocephalus was defined as: 1) modified frontal horn index greater than 33%, and 2) the presence of Gudeman CT criteria. Hygromas were diagnosed based on subdural fluid collection and classified according to location of the craniectomy. Hydrocephalus was diagnosed in 20 patients (27.4%). After uni- and multivariate analysis, the presence of interhemispheric hygromas (IHHs) was the only independent prognostic factor for development of posttraumatic hydrocephalus (p<0.0001). More than 80% of patients with IHHs developed hydrocephalus within the first 50 days of undergoing DC. In all cases the presence of hygromas preceded the diagnosis of hydrocephalus. The IHH predicts the development of hydrocephalus after DC with 94% sensitivity and 96% specificity. The presence of an IHH showed an area under the receiver-operator characteristic of 0.951 (95% CI 0.87-1.00; p<0.0001). Hydrocephalus was observed in 27.4% of the patients with severe traumatic brain injury who required DC. The presence of IHHs was a predictive radiological sign of hydrocephalus development within the first 6 months of DC in patients with severe head injury.


Calcium imbalance in hypoparathyroidism.
Extracellular fluid calcium is a tightly controlled variable. Hypoparathyroid state may result in profound calcium imbalance and moderate to severe hypocalcaemia. During 1974-89, 108 cases of hypoparathyroidism (97 post-surgical and 11 idiopathic) were seen. In the post-thyroidectomy group, 83 cases (85%) presented with acute transient hypocalcaemia with spontaneous recovery within 7-10 days. Chronic hypoparathyroidism was seen in 25 cases (14 post-surgical and 11 idiopathic). Convulsions resembling epileptic fits were seen in 9 cases (36%). Pseudopapilloedema was seen in three cases presenting with fits. The administration of phenobarbitone and dilantin aggravated convulsions in 9 patients. The other manifestations were psychiatric illness, cataract and calcification of basal ganglion. Biochemical findings included persistent hypocalcaemia with normal or raised serum phosphorus and lowered daily urinary excretion of calcium. Twenty three of 25 chronic hypoparathyroid cases were treated with vitamin D3 (1-3 mg/day) and calcium supplements (600-1000 mg/day)while 1 alfa-calcidol or calcitriol was used in two patients. Four patients receiving treatment with vitamin D3 developed transient hypercalcaemia with raised plasma levels of 25 hydroxy-vitamin D3. They responded to a reduction in dosage of vitamin D3. One patient was later changed over to 1-alfa-calcidol and another to calcitriol.


The mouse mammary tumour virus promoter positioned on a tetramer of histones H3 and H4 binds nuclear factor 1 and OTF1.
Modulation of eukaryotic gene expression is influenced by the organization of regulatory DNA-elements in chromatin. The mouse mammary tumor virus (MMTV) promoter exhibits regularly positioned nucleosomes that reduce the accessibility of the binding sites for sequence-specific transcription factors, in particular nuclear factor (NF1). Hormonal induction of the MMTV promoter is accompanied by remodeling of the nucleosomal structure, but the biochemical nature of these structural changes is unknown. Using recombinant histones, we have now assembled the MMTV promoter in particles containing either an octamer of the histones H3, H4, H2A and H2B or a tetramer of histones H3 and H4, and have compared the two particles in terms of structure, positioning, and exclusion of transcription factors. Using site-directed hydroxy radicals to map histone locations, two main nucleosome positions are found with dyads at position -107 and at -127. The same two main positions are found for particles containing only the H3/H4 tetramer, showing that the absence of H2A/H2B dimers does not alter positioning. The rotational orientation of the DNA double helix in both types of particles is essentially identical. However, the ends of the nucleosomal DNA as well as its central region are more accessible to cleavage reagents in the tetramer particle than in the octamer particle. In agreement with these structural features, the transcription factors NF1 and OTF1 were able to bind to their cognate sites on the tetramer particle, while they could not gain access to the same sites on the surface of the octamer particle. The DNase I digestion pattern of octamers treated with partially purified SWI/SNF complex from HeLa cells in the presence of ATP is indistinguishable from that of tetramer particles, suggesting that the SWI/SNF complex promotes ATP-dependent remodeling of the octamer particle but not of tetramer particles. These results are compatible with a hormone-induced removal of histone H2A/H2B during MMTV induction.


The specious interaction of time and numerosity perception.
Magnitude information is essential to create a representation of the external environment and successfully interact with it. Duration and numerosity, for example, can shape our predictions and bias each other (i.e. the greater the number of people queuing, the longer we expect to wait). While these biases suggest the existence of a generalized magnitude system, asymmetric effects (i.e. numerosity affecting duration but not vice versa) challenged this idea. Here, we propose that such asymmetric integration depends on the stimuli used and the neural processing dynamics they entail. Across multiple behavioural experiments employing different stimulus presentation displays (static versus dynamic) and experimental manipulations known to bias numerosity and duration perceptions (i.e. connectedness and multisensory integration), we show that the integration between numerosity and time can be symmetrical if the stimuli entail a similar neural time-course and numerosity unfolds over time. Overall, these findings support the idea of a generalized magnitude system, but also highlight the role of early sensory processing in magnitude representation and integration.


Prognostic impact of tumour-specific HMG-CoA reductase expression in primary breast cancer.
We have previously reported that tumour-specific expression of the rate-limiting enzyme, 3-hydroxy-3-methylglutharyl-coenzyme A reductase (HMG-CoAR), in the mevalonate pathway is associated with more favourable tumour parameters in breast cancer. In the present study, we examined the prognostic value of HMG-CoAR expression in a large cohort of primary breast cancer patients with long-term follow up. The expression of HMG-CoAR was assessed by immunohistochemistry on tissue microarrays with tumour specimens from 498 consecutive cases of breast cancer with a median follow-up of 128 months. Kaplan Meier analysis and Cox proportional hazards modelling were used to estimate the rate of recurrence-free survival (RFS) and breast cancer specific survival (BCSS). In line with our previous findings, tumour-specific HMG-CoAR expression was associated with low grade (p < 0.001), small size (p = 0.007), oestrogen receptor (ER) positive (p = 0.01), low Ki-67 (p = 0.02) tumours. Patients with tumours expressing HMG-CoAR had a significantly prolonged RFS, even when adjusted for established prognostic factors (relative risk [RR] = 0.60, 95% confidence interval [CI] 0.40 to 0.92; p = 0.02). In ER-negative tumours, however, there was a trend, that was not significantly significant, towards a shorter RFS in HMG-CoAR expressing tumours. HMG-CoAR expression is an independent predictor of a prolonged RFS in primary breast cancer. This may, however, not be true for ER-negative tumours. Further studies are needed to shed light on the value of HMG-CoAR expression as a surrogate marker of response to statin treatment, especially with respect to hormone receptor status.


Evaluation of the Incidence of New-Onset Atrial Fibrillation After Aortic Valve Replacement.
Data on the burden of new-onset atrial fibrillation after transcatheter aortic valve implantation (TAVI) and surgical aortic valve replacement (AVR) is limited mostly to small series or post hoc analyses of clinical trials. To evaluate the incidence of new-onset atrial fibrillation and assess the incidence of in-hospital mortality associated with new-onset atrial fibrillation after TAVI and AVR. In this population-based observational study using the National Inpatient Sample and a validation cohort from the New York state inpatient database, the National Inpatient Sample was queried from January 1, 2012, to September 30, 2015, and the New York state inpatient database was queried from January 1, 2012, to December 31, 2014. Hospitalizations of adults undergoing TAVI or isolated AVR were examined. The incidence of in-hospital mortality across groups with new-onset atrial fibrillation was assessed in the National Inpatient Sample cohort using multivariable logistic regression modeling. Statistical analysis was conducted from August 20, 2018, to March 19, 2019. The primary outcome was the occurrence of new-onset atrial fibrillation, which was identified by excluding hospitalizations in which atrial fibrillation was present on admission. The secondary outcome was in-hospital mortality in TAVI and AVR hospitalizations with and without new-onset atrial fibrillation. A total of 48 715 TAVI hospitalizations (47.4% women and 52.6% men; mean [SD] age, 81.3 [8.1] years; 82.3% white) and 122 765 AVR hospitalizations (39.0% women and 61.0% men; mean [SD] age, 67.8 [12.0] years; 78.0% white) were identified. New-onset atrial fibrillation occurred in 50.4% of TAVI hospitalizations and 50.1% of AVR hospitalizations. In the multivariable-adjusted model, TAVI and AVR hospitalizations with new-onset atrial fibrillation had higher odds of in-hospital mortality compared with hospitalizations without new-onset atrial fibrillation (TAVI: odds ratio, 1.57; 95% CI, 1.21-2.04; and AVR: odds ratio, 1.36; 95% CI, 1.08-1.70). The results were then confirmed with the New York state inpatient database, which contains a present on arrival indicator. The incidence of new-onset atrial fibrillation was 14.1% (244 of 1736 hospitalizations) after TAVI and 30.6% (1573 of 5141 hospitalizations) after AVR in the New York state inpatient database. In this large nationwide study, a substantial burden of new-onset atrial fibrillation was observed after TAVI and AVR. The incidence of new-onset atrial fibrillation was higher after AVR than after TAVI in a patient-level state inpatient database.


Crystallinity of the double layer of cadmium arachidate films at the water surface.
A crystalline counterionic layer at the interface between an electrolyte solution and a charged layer of insoluble amphiphilic molecules was observed with grazing incidence synchrotron x-ray diffraction. Uncompressed arachidic films spread over 10(-3) molar cadmium chloride solution (pH 8.8) spontaneously form crystalline clusters with coherence lengths of approximately 1000 angstroms at 9 degrees C. Ten distinct diffraction peaks were observed, seven of which were attributed to scattering only from a crystalline Cd(2+) layer and the other three to scattering primarily from the arachidate layer. The reflections from the Cd(2+) layer were indexed according to a 2 x 3 supercell of the arachidate lattice with three Cd(2+) ions per cadmium unit cell.


Management of advanced or recurrent cervical cancer: chemotherapy and beyond.
Metastatic or recurrent cervical cancer has a survival duration of approximately 12 months. Thus, we review the outcomes of chemotherapy and/or novel agents for women who present in this situation. Included were studies in women with advanced or recurrent cervical cancer where at least response rate, survival or toxicity were reported. Platinum alone chemotherapy at 50 mg/m(2) is superior to other single-agent platinum or nonplatinum regimens in terms of efficacy and toxicity profile. Platinum with either paclitaxel, gemcitibine, topotecan or vinorelbine has equivalent efficacy but differ with respect to toxicity profile and convenience of treatment regimen. Bevacizumab in addition to combination chemotherapy provides a duration of survival that is 3.7 months longer than that provided by combination chemotherapy alone. EGF receptor (EGFR) tyrosine kinase inhibitor agents did not provide promising efficacy. Bevacizumab in addition to combination chemotherapy provides superior efficacy with acceptable toxicity. Evaluation of other novel targeted antiangiogenic agents, either alone or in combination with chemotherapy is ongoing.


Pulmonary delivery of an erythropoietin Fc fusion protein in non-human primates through an immunoglobulin transport pathway.
Administration of therapeutic proteins by methods other than injection is limited, in part, by inefficient penetration of epithelial barriers. Therefore, unique approaches to breaching these barriers are needed. The neonatal constant region fragment (Fc) receptor (FcRn), which is responsible for IgG transport across the intestinal epithelium in newborn rodents, is expressed in epithelial cells in adult humans and non-human primates. Here we show that FcRn-mediated transport is functional in the lung of non-human primates and that this transport system can be used to deliver erythropoietin (Epo) when it is conjugated to the Fc domain of IgG1. FcRn-dependent absorption was more efficient when the EpoFc fusion protein was deposited predominantly in the upper and central airways of the lung, where epithelial expression of FcRn was most prominently detected. To optimize fusion protein absorption in the lung, we created a recombinant "monomeric-Epo" Fc fusion protein comprised of a single molecule of Epo conjugated to a dimeric Fc. This fusion protein exhibited enhanced pharmacokinetic and pharmacodynamic properties. The bioavailability of the EpoFc monomer when delivered through the lung was approximately equal to that reported for unconjugated Epo delivered s.c. in humans. These studies show that FcRn can be harnessed to noninvasively deliver bioactive proteins into the systemic circulation in therapeutic quantities.


Elevated CO2 and salinity are responsible for phenolics-enrichment in two differently pigmented lettuces.
Both salt stress and high CO2 level, besides influencing secondary metabolism, can affect oxidative status of plants mainly acting in an opposite way with salinity provoking oxidative stress and elevated CO2 alleviating it. The aim of the present work was to study the changes in the composition of phenolic acids and flavonoids as well as in the antioxidant activity in two differently pigmented lettuce cvs (green or red leaf) when submitted to salinity (200 mM NaCl) or elevated CO2 (700 ppm) or to their combination in order to evaluate how a future global change can affect lettuce quality. Following treatments, the red cv. always maintained higher levels of antioxidant secondary metabolites as well as antioxidant activity, proving to be more responsive to altered environmental conditions than the green one. Overall, these results suggest that the application of moderate salinity or elevated CO2, alone or in combination, can induce the production of some phenolics that increase the health benefits of lettuce. In particular, moderate salinity was able to induce the synthesis of the flavonoids quercetin, quercetin-3-O-glucoside, quercetin-3-O-glucuronide and quercitrin. Phenolics-enrichment as well as a higher antioxidant capacity were also observed under high CO2 with the red lettuce accumulating cyanidin, free chlorogenic acid, conjugated caffeic and ferulic acid as well as quercetin, quercetin-3-O-glucoside, quercetin-3-O-glucuronide, luteolin-7-O-glucoside, rutin, quercitrin and kaempferol. When salinity was present in combination with elevated CO2, reduction in yield was prevented and a higher presence of phenolic compounds, in particular luteolin, was observed compared to salinity alone.


Optimal control of hepatitis C antiviral treatment programme delivery for prevention amongst a population of injecting drug users.
In most developed countries, HCV is primarily transmitted by injecting drug users (IDUs). HCV antiviral treatment is effective, and deemed cost-effective for those with no re-infection risk. However, few active IDUs are currently treated. Previous modelling studies have shown antiviral treatment for active IDUs could reduce HCV prevalence, and there is emerging interest in developing targeted IDU treatment programmes. However, the optimal timing and scale-up of treatment is unknown, given the real-world constraints commonly existing for health programmes. We explore how the optimal programme is affected by a variety of policy objectives, budget constraints, and prevalence settings. We develop a model of HCV transmission and treatment amongst active IDUs, determine the optimal treatment programme strategy over 10 years for two baseline chronic HCV prevalence scenarios (30% and 45%), a range of maximum annual budgets (£50,000-300,000 per 1,000 IDUs), and a variety of objectives: minimising health service costs and health utility losses; minimising prevalence at 10 years; minimising health service costs and health utility losses with a final time prevalence target; minimising health service costs with a final time prevalence target but neglecting health utility losses. The largest programme allowed for a given budget is the programme which minimises both prevalence at 10 years, and HCV health utility loss and heath service costs, with higher budgets resulting in greater cost-effectiveness (measured by cost per QALY gained compared to no treatment). However, if the objective is to achieve a 20% relative prevalence reduction at 10 years, while minimising both health service costs and losses in health utility, the optimal treatment strategy is an immediate expansion of coverage over 5-8 years, and is less cost-effective. By contrast, if the objective is only to minimise costs to the health service while attaining the 20% prevalence reduction, the programme is deferred until the final years of the decade, and is the least cost-effective of the scenarios.


The magnetic compass mechanisms of birds and rodents are based on different physical principles.
Recently, oscillating magnetic fields in the MHz-range were introduced as a useful diagnostic tool to identify the mechanism underlying magnetoreception. The effect of very weak high-frequency fields on the orientation of migratory birds indicates that the avian magnetic compass is based on a radical pair mechanism. To analyse the nature of the magnetic compass of mammals, we tested rodents, Ansell's mole-rats, using their tendency to build their nests in the southern part of the arena as a criterion whether or not they could orient. In contrast to birds, their orientation was not disrupted when a broad-band field of 0.1-10MHz of 85nT or a 1.315MHz field of 480nT was added to the static geomagnetic field of 46000nT. Even increasing the intensity of the 1.315MHz field (Zeeman frequency in the local geomagnetic field) to 4800nT, more than a tenth of the static field, the mole-rats remained unaffected and continued to build their nests in the south. These results indicate that in contrast to that of birds, their magnetic compass does not involve radical pair processes; it seems to be based on a fundamentally different principle, which probably involves magnetite.


Ziprasidone Augmentation of Escitalopram for Major Depressive Disorder: Efficacy Results From a Randomized, Double-Blind, Placebo-Controlled Study.
The authors sought to test the efficacy of adjunctive ziprasidone in adults with nonpsychotic unipolar major depression experiencing persistent symptoms after 8 weeks of open-label treatment with escitalopram. This was an 8-week, randomized, double-blind, parallel-group, placebo-controlled trial conducted at three academic medical centers. Participants were 139 outpatients with persistent symptoms of major depression after an 8-week open-label trial of escitalopram (phase 1), randomly assigned in a 1:1 ratio to receive adjunctive ziprasidone (escitalopram plus ziprasidone, N=71) or adjunctive placebo (escitalopram plus placebo, N=68), with 8 weekly follow-up assessments. The primary outcome measure was clinical response, defined as a reduction of at least 50% in score on the 17-item Hamilton Depression Rating Scale (HAM-D). The Hamilton Anxiety Rating scale (HAM-A) and Visual Analog Scale for Pain were defined a priori as key secondary outcome measures. Rates of clinical response (35.2% compared with 20.5%) and mean improvement in HAM-D total scores (-6.4 [SD=6.4] compared with -3.3 [SD=6.2]) were significantly greater for the escitalopram plus ziprasidone group. Several secondary measures of antidepressant efficacy also favored adjunctive ziprasidone. The escitalopram plus ziprasidone group also showed significantly greater improvement on HAM-A score but not on Visual Analog Scale for Pain score. Ten (14%) patients in the escitalopram plus ziprasidone group discontinued treatment because of intolerance, compared with none in the escitalopram plus placebo group. Ziprasidone as an adjunct to escitalopram demonstrated antidepressant efficacy in adult patients with major depressive disorder experiencing persistent symptoms after 8 weeks of open-label treatment with escitalopram.


Triphenyltin chloride induces glucose intolerance by the suppression of insulin release from hamster pancreatic beta-cells.
We performed an intravenous glucose tolerance test (IVGTT) after the administration of triphenyltin chloride (TPTCl-Ad) in hamsters in order to confirm the presence of glucose intolerance and to clarify the pathogenesis of TPTCl-induced glucose intolerance. On the 1st, 2nd, 3rd, 4th and 7th days after TPTCl-Ad or the administration of sesame oil alone as a control, glucose was injected at a dose of 0.05 g glucose/100 g B.W., and then PG, IRI and TG were measured on 0, 1, 2, 3, 5 and 10 min after IVGTT. The TPT concentration in the pancreas was measured by gaschromatography and the morphological findings with a transmission electron microscope were compared between the TPTCl-Ad and the control hamsters. FPG on the 1st and the 2nd days after TPTCl-Ad were significantly higher than those in control, but those on the 4th and the 7th day recovered up to the control level. In contrast, basal IRI levels showed reciprocal results compared to the FPG levels. delta IRI/ delta PG on the 1st day after TPTCl-Ad was significantly reduced compared to the control. Fasting TG on the 1st day after TPTCl-Ad was much higher than the control. TPT-concentration on the 1st day after TPTCl-Ad showed peak values and its concentration gradually decreased. Electron microscopic findings in the pancreas after TPTCl-Ad indicated neither destruction nor lymphocyte infiltration of the pancreas. The present data suggest that the administration of TPTCl in hamsters induces a functional transient damage on islet cells but not a morphological disorder, which shows an essentially different nature from the change in the pancreas induced by viral infection or by a large amount of streptozotocin.


Structure of RNA 3'-phosphate cyclase bound to substrate RNA.
RNA 3'-phosphate cyclase (RtcA) catalyzes the ATP-dependent cyclization of a 3'-phosphate to form a 2',3'-cyclic phosphate at RNA termini. Cyclization proceeds through RtcA-AMP and RNA(3')pp(5')A covalent intermediates, which are analogous to intermediates formed during catalysis by the tRNA ligase RtcB. Here we present a crystal structure of Pyrococcus horikoshii RtcA in complex with a 3'-phosphate terminated RNA and adenosine in the AMP-binding pocket. Our data reveal that RtcA recognizes substrate RNA by ensuring that the terminal 3'-phosphate makes a large contribution to RNA binding. Furthermore, the RNA 3'-phosphate is poised for in-line attack on the P-N bond that links the phosphorous atom of AMP to N(ε) of His307. Thus, we provide the first insights into RNA 3'-phosphate termini recognition and the mechanism of 3'-phosphate activation by an Rtc enzyme.


Relationship Between Arterial Access and Outcomes in ST-Elevation Myocardial Infarction With a Pharmacoinvasive Versus Primary Percutaneous Coronary Intervention Strategy: Insights From the STrategic Reperfusion Early After Myocardial Infarction (STREAM) Study.
The effectiveness of radial access (RA) in ST-elevation myocardial infarction (STEMI) has been predominantly established in primary percutaneous coronary intervention (pPCI) with limited exploration of this issue in the early postfibrinolytic patient. The purpose of this study was to compare the effectiveness and safety of RA versus femoral (FA) access in STEMI undergoing either a pharmacoinvasive (PI) strategy or pPCI. Within STrategic Reperfusion Early After Myocardial Infarction (STREAM), we evaluated the relationship between arterial access site and primary outcome (30-day composite of death, shock, congestive heart failure, or reinfarction) and major bleeding according to the treatment strategy received. A total of 1820 STEMI patients were included: 895 PI (49.2%; rescue PCI [n=379; 42.3%], scheduled PCI [n=516; 57.7%]) and 925 pPCI (50.8%). Irrespective of treatment strategy, there was comparable utilization of either access site (FA: PI 53.4% and pPCI 57.6%). FA STEMI patients were younger, had lower presenting systolic blood pressure, lesser Thrombolysis In Myocardial Infarction risk, and more ∑ST-elevation at baseline. The primary composite endpoint occurred in 8.9% RA versus 15.7% FA patients (P<0.001). On multivariable analysis, this benefit on the primary composite outcome favoring RA persisted (adjusted odds ratio [OR], 0.59; 95% CI, 0.44-0.78; P<0.001) and was evident in both pPCI (adjusted OR, 0.63; 95% CI, 0.43-0.92) and PI cohorts (adjusted OR, 0.57 95% CI, 0.37-0.86; P interaction=0.730). There was no difference in nonintracranial major bleeding with either access group (RA vs FA, 5.2% vs 6.0%; P=0.489). Regardless of the application of a PI or pPCI strategy, RA was associated with improved clinical outcomes, supporting current STEMI evidence in favor of RA in PCI. URL: https://www.clinicaltrials.gov/. Unique identifier: NCT00623623.


Competing causes of death and medical comorbidities among patients with human papillomavirus-positive vs human papillomavirus-negative oropharyngeal carcinoma and impact on adherence to radiotherapy.
Survival of patients with head and neck cancer can be affected by competing causes of mortality, as well as comorbidities that result in radiation treatment interruptions. To discern how differences in preexisting medical and psychosocial comorbidities potentially influence adherence to radiation therapy according to human papillomavirus (HPV) status. Retrospective analysis at a comprehensive cancer center of 162 consecutive patients with locally advanced squamous cell carcinoma of the oropharynx treated with primary chemoradiation (n = 95) or primary surgery followed by adjuvant radiation (n = 67). Immunostaining for p16 was used to determine HPV status. Difference in alcohol, tobacco, and marijuana use was compared between patients with HPV-positive and HPV-negative tumors, as well as the prevalence of the following comorbidities: diabetes mellitus, chronic obstructive pulmonary disease (COPD), anxiety disorder, and major depression. The number of total missed treatment days was analyzed as both a continuous and categorical variable. Rates of self-reported heavy alcohol use (47% vs 16%; P = .02) and any marijuana use (47% vs 23%; P = .003) were significantly higher among HPV-negative patients. Fifty-four percent of HPV-positive patients self-identified as never smokers, compared with only 12% of HPV-negative patients (P < .001). HPV-negative patients had more missed treatment days (mean, 2.8 vs 1.7 days; P = .02), as well as an increased rate of at least 5 missed days (24% vs 10%; P = .04), and higher prevalences of COPD (12% vs 7%; P = .37) and anxiety disorder (12% vs 6%; P = .35). Pronounced differences exist in lifestyle habits between patients with HPV-negative and HPV-positive oropharyngeal cancer at diagnosis. These differences, as well as those of medical and psychosocial burden, may contribute to observed discrepancies in treatment adherence and need to be considered in outcomes reporting and clinical trial design.


Anastomotic Leak Does Not Impact Oncologic Outcomes After Preoperative Chemoradiotherapy and Resection for Rectal Cancer.
The aim of this study was to evaluate the relationship of anastomotic leakage, local recurrence, and overall survival in rectal cancer patients treated with preoperative chemoradiotherapy (CRT) and curative resection. Little is known about the association between anastomotic leakage and oncologic outcomes after preoperative CRT. A total of 698 consecutive primary rectal cancer patients after preoperative CRT between April 19, 2000, and December 27, 2013, were retrospectively reviewed. Forty-seven patients who had anastomotic leakage were compared with 651 patients who had no anastomotic leakage. Of 698 patients, 47 (6.7%) patients had anastomotic leakage. Among these 47 patients, 39 (83.0%) had grade C leak that required urgent operation, while 8 (17.0%) had grade B leak that was managed expectantly or by percutaneous drainage. The median follow-up period was 47.6 months (range, 27.1 to 68.9 months). One hundred twenty (17.2%) recurrences were identified among all patients. The median overall disease-free survival was 43 months (range, 22.4 to 66.7 months). Five-year disease-free survival did not differ significantly between the 2 groups (80.5% vs 80.4%, P = 0.839). Five-year local recurrence-free survival did not differ significantly either between the 2 groups (93.7% vs 94.9%, P = 0.653). Five-year overall survival rates of patients with or without leakage were 90.9% and 86.3%, respectively (P = 0.242). Five-year cancer-specific survival rates of patients with or without leakage were 92.2% and 86.3%, respectively (P = 0.248). After preoperative CRT, an anastomotic leak is not associated with a significant increase in local recurrence or long-term survival in rectal cancer.


Combined bi-occipital suboccipital transsinus transtentorial approach for resection of a pineal region falcotentorial meningioma: operative video and technical nuances.
Large deep-seated meningiomas of the falcotentorial region present a formidable surgical challenge. In this operative video, the author demonstrates the combined bi-occipital suboccipital transsinus transtentorial approach for microsurgical resection of a large falcotentorial meningioma. This approach involves division of the less dominant transverse sinus after assessment of the venous pressure before and after clipping of the sinus with continuous neurophysiologic monitoring. Mild retraction of the occipital lobe and cerebellum results in a wide supra- and infratentorial exposure of extensive pineal region tumors. This video atlas demonstrates the operative technique and surgical nuances, including patient positioning, supra- and infratentorial craniotomy, transsinus transtentorial incision, and tumor removal with preservation of the vein of Galen complex. In summary, the combined bi-occipital suboccipital transsinus transtentorial approach provides a wide supra- and infratentorial surgical corridor for removal of select falcotentorial meningiomas. The video can be found here: https://youtu.be/3aD8h2uwBAo .


Brain architecture in the terrestrial hermit crab Coenobita clypeatus (Anomura, Coenobitidae), a crustacean with a good aerial sense of smell.
During the evolutionary radiation of Crustacea, several lineages in this taxon convergently succeeded in meeting the physiological challenges connected to establishing a fully terrestrial life style. These physiological adaptations include the need for sensory organs of terrestrial species to function in air rather than in water. Previous behavioral and neuroethological studies have provided solid evidence that the land hermit crabs (Coenobitidae, Anomura) are a group of crustaceans that have evolved a good sense of aerial olfaction during the conquest of land. We wanted to study the central olfactory processing areas in the brains of these organisms and to that end analyzed the brain of Coenobita clypeatus (Herbst, 1791; Anomura, Coenobitidae), a fully terrestrial tropical hermit crab, by immunohistochemistry against synaptic proteins, serotonin, FMRFamide-related peptides, and glutamine synthetase. The primary olfactory centers in this species dominate the brain and are composed of many elongate olfactory glomeruli. The secondary olfactory centers that receive an input from olfactory projection neurons are almost equally large as the olfactory lobes and are organized into parallel neuropil lamellae. The architecture of the optic neuropils and those areas associated with antenna two suggest that C. clypeatus has visual and mechanosensory skills that are comparable to those of marine Crustacea. In parallel to previous behavioral findings of a good sense of aerial olfaction in C. clypeatus, our results indicate that in fact their central olfactory pathway is most prominent, indicating that olfaction is a major sensory modality that these brains process. Interestingly, the secondary olfactory neuropils of insects, the mushroom bodies, also display a layered structure (vertical and medial lobes), superficially similar to the lamellae in the secondary olfactory centers of C. clypeatus. More detailed analyses with additional markers will be necessary to explore the question if these similarities have evolved convergently with the establishment of superb aerial olfactory abilities or if this design goes back to a shared principle in the common ancestor of Crustacea and Hexapoda.


Sequences of purchases in credit card data reveal lifestyles in urban populations.
Zipf-like distributions characterize a wide set of phenomena in physics, biology, economics, and social sciences. In human activities, Zipf's law describes, for example, the frequency of appearance of words in a text or the purchase types in shopping patterns. In the latter, the uneven distribution of transaction types is bound with the temporal sequences of purchases of individual choices. In this work, we define a framework using a text compression technique on the sequences of credit card purchases to detect ubiquitous patterns of collective behavior. Clustering the consumers by their similarity in purchase sequences, we detect five consumer groups. Remarkably, post checking, individuals in each group are also similar in their age, total expenditure, gender, and the diversity of their social and mobility networks extracted from their mobile phone records. By properly deconstructing transaction data with Zipf-like distributions, this method uncovers sets of significant sequences that reveal insights on collective human behavior.


Kineobactrum sediminis gen. nov., sp. nov., isolated from marine sediment.
A novel Gram-stain-negative, rod-shaped marine bacterium, designated strain F02T, was isolated from a marine saltern in Weihai, PR China. The cells of strain F02T were approximately 0.8-1.0×3.0-4.0 µm and motile by means of a polar flagellum. Strain F02T grew optimally at 33-35 °C, pH 7.5 and in the presence of 3.0 % (w/v) NaCl. Strain F02T showed oxidase- and catalase-positive activities. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain F02T belonged to the family Halieaceae and exhibited 16S rRNA gene sequence similarities of 96.6 and 96.4 % to the type strains of Chromatocurvus halotolerans and Parahaliea mediterranea, respectively. The major cellular fatty acids of strain F02T were C18 : 1ω7c, C16 : 1ω7c, C15 : 0 and C18 : 1ω9c. The major polar lipids of strain F02T were phosphatidylglycerol, phosphatidylethanolamine and one unidentified aminolipid. Strain F02T contained Q-8 as the sole respiratory quinone. The genomic DNA G+C content was 58.4 mol%. The genome sequences of strain F02T and Chromatocurvus halotolerans DSM 23344T had an OrthoANI value of 70.5 %, and the average amino acid identity value between the two genomes was 62.6 %. The sequence similarity value between the rpoB genes of strain F02T and Chromatocurvus halotolerans DSM 23344T was 79.5 %. On the basis of polyphasic analysis, strain F02T represents a novel species in a new genus, for which the name Kineobactrum sediminis gen. nov., sp. nov. is proposed. The type strain is F02T (=KCTC 52616T=MCCC 1H00224T).


[Molecular cloning of the structural genes of the Escherichia coli deo-operon in plasmid RSF2124].
A specialized phage lambda ddeo carrying the deo operon of Escherichia coli is analyzed by exposing the DNA to the specific restriction endonucleases EcoRI and BamHI. Using the lambda ddeo DNA fragment, obtained by digestion with BamHI and plasmid RSF2124 as a vehicle, the hybrid plasmid pAM1 carrying all the genes of the deo operon is constructed and cloned in E. coli cells. It is shown that the activity of thymidine phosphorylase in the strain AM061, which contains hybrid plasmid pAM1 is 30-fold greater than that in strains of E. coli with chromosomal localization of the deo operon.


Structural features of the regulatory ACT domain of phenylalanine hydroxylase.
Phenylalanine hydroxylase (PAH) catalyzes the conversion of L-Phe to L-Tyr. Defects in PAH activity, caused by mutations in the human gene, result in the autosomal recessively inherited disease hyperphenylalaninemia. PAH activity is regulated by multiple factors, including phosphorylation and ligand binding. In particular, PAH displays positive cooperativity for L-Phe, which is proposed to bind the enzyme on an allosteric site in the N-terminal regulatory domain (RD), also classified as an ACT domain. This domain is found in several proteins and is able to bind amino acids. We used molecular dynamics simulations to obtain dynamical and structural insights into the isolated RD of PAH. Here we show that the principal motions involve conformational changes leading from an initial open to a final closed domain structure. The global intrinsic motions of the RD are correlated with exposure to solvent of a hydrophobic surface, which corresponds to the ligand binding-site of the ACT domain. Our results strongly suggest a relationship between the Phe-binding function and the overall dynamic behaviour of the enzyme. This relationship may be affected by structure-disturbing mutations. To elucidate the functional implications of the mutations, we investigated the structural effects on the dynamics of the human RD PAH induced by six missense hyperphenylalaninemia-causing mutations, namely p.G46S, p.F39C, p.F39L, p.I65S, p.I65T and p.I65V. These studies showed that the alterations in RD hydrophobic interactions induced by missense mutations could affect the functionality of the whole enzyme.


ASA class is associated with early revision and reoperation after total hip arthroplasty: an analysis of the Geneva and Swedish Hip Arthroplasty Registries.
Background and purpose - Data from several joint replacement registries suggest that the rate of early revision surgery after primary total hip arthroplasty (THA) is increasing. The ASA class, now widely recorded in arthroplasty registries, may predict early revision. We investigated the influence of ASA class on the risk of revision and other reoperation within 3 months and within 5 years of primary THA. Patients and methods - We used data from the Geneva and Swedish Hip Arthroplasty Registries, on primary elective THAs performed in 1996-2016 and 2008-2016, respectively. 5,319 and 122,241 THAs were included, respectively. Outcomes were all-cause revision and other reoperations evaluated using Kaplan-Meier survival and Cox regression analyses. Results - Within 3 months after surgery, higher ASA class was associated with greater risk of revision and other reoperation. 3-month cumulative incidences of revision by ASA class I, II, and III-IV respectively, were 0.6%, 0.7%, and 2.3% in Geneva and 0.5%, 0.8%, and 1.6% in Sweden. 3-month cumulative incidences of other reoperation were 0.4%, 0.7%, and 0.9% in Geneva and 0.2%, 0.4%, and 0.7% in Sweden. Adjusted hazard ratios (ASA III-IV vs. I) for revision within 3 months were 2.7 (95% CI 1.2-5.9) in Geneva and 3.3 (CI 2.6-4.0) in Sweden. Interpretation - Assessment of ASA class of patients prior to THA will facilitate risk stratification. Targeted risk-reduction strategies may be appropriate during the very early postoperative period for patients identified as at higher risk. Systematically recording ASA class in arthroplasty registries will permit risk adjustment and facilitate comparison of revision rates internationally.


[Femtosecond laser assisted cataract surgery in a complicated cataract patient with reverse implantable collamer len: a case report].
The patient is a 33-year-old female who, 11 years ago, underwent bilateral posterior chamber phakic intraocular lens (pIOL) implantation due to myopia. She presented with a 2-year history of declining vision in her right eye and sought medical attention. She received femtosecond laser-assisted cataract surgery combined with pIOL extraction. Anterior segment optical coherence tomography and ultrasound biomicroscopy both showed an inverted pIOL in the right eye. Good visual results were achieved, and there were no complications during the six-month follow-up.


Magnaporthe oryzae isolates causing gray leaf spot of perennial ryegrass possess a functional copy of the AVR1-CO39 avirulence gene.
SUMMARY Gray leaf spot of perennial ryegrass (Lolium perenne) is a severe foliar disease caused by the ascomycete fungus Magnaporthe oryzae (formerly known as Magnaporthe grisea). Control of gray leaf spot is completely dependent on the use of fungicides because currently available perennial ryegrass cultivars lack genetic resistance to this disease. M. oryzae isolates from perennial ryegrass (prg) were unable to cause disease on rice cultivars CO39 and 51583, and instead triggered a hypersensitive response. Southern hybridization analysis of DNA from over 50 gray leaf spot isolates revealed that all of them contain sequences corresponding to AVR1-CO39, a host specificity gene that confers avirulence to rice cultivar CO39, which carries the corresponding resistance gene Pi-CO39(t). There was also an almost complete lack of restriction site polymorphism at the avirulence locus. Cloning and sequencing of the AVR1-CO39 gene (AVR1-CO39(Lp)) from 16 different gray leaf spot isolates revealed just two point mutations, both of which were located upstream of the predicted open reading frame. When an AVR1-CO39(Lp) gene copy was transferred into ML33, a rice pathogenic isolate that is highly virulent to rice cultivar CO39, the transformants were unable to cause disease on CO39 but retained their virulence to 51583, a rice cultivar that lacks Pi-CO39(t). These data demonstrate that the AVR1-CO39 gene in the gray leaf spot pathogens is functional, and suggest that interaction of AVR1-CO39(Lp) and Pi-CO39(t) is responsible, at least in part, for the host specificity expressed on CO39. This indicates that it may be possible to use the Pi-CO39(t) resistance gene as part of a transgenic strategy to complement the current deficiency of gray leaf spot resistance in prg. Furthermore, our data indicate that, if Pi-CO39(t) can function in prg, the resistance provided should be broadly effective against a large proportion of the gray leaf spot pathogen population.


A novel feature enhancement and semantic segmentation scheme for identifying low-contrast ocean oil spills.
The oil spill accidents on the sea surface pose a severe threat to the marine environment and human health. This paper proposes a novel Semantic Segmentation Network (SSN) for processing oil spill images so that low-contrast oil spills on the sea surface can be accurately identified. After the detection accuracy and real-time performance of the current SSNs are compared, the basic network architecture of DeeplabV3+ based target detection is analyzed. The standard convolution is replaced by the Omni-dimensional Dynamic Convolution (ODConv) in the Ghost Module Depth-Wise separable Convolution (DWConv) to further enhance the feature extraction ability of the network. Furthermore, a new DeeplabV3+ based network with ODGhostNetV2 is constructed as the main feature extraction module, and an Adaptive Triplet Attention (ATA) module is deployed in the encoder and decoder at the same time. This not only improves the richness of semantic features but also increases the following receptive fields of the network model. ATA integrates the Adaptively Spatial Feature Fusion (ASFF) module to optimize the weight assignment problem in the feature map fusion process. The ablation experiments are conducted to verify the proposed network which show high accuracy and good real-time performance for the oil spill detection.


The flagellum-mitogen-activated protein kinase connection in Trypanosomatids: a key sensory role in parasite signalling and development?
Trypanosomatid parasites are the causative agents of severe human diseases such as sleeping sickness, Chagas disease and leishmaniases. These microorganisms are transmitted via different insect vectors and hence are confronted to changing environments during their infectious cycle in which they activate specific and complex patterns of differentiation. Several studies in Trypanosoma brucei and in different subspecies of Leishmania have shed light on the role of mitogen-activated protein (MAP) kinases in these processes. Surprisingly, several MAP kinases turned out to be involved in the control of flagellum length in the promastigote stage of Leishmania. Recently, a sensory function has been recognized for cilia and flagella in unicellular and multicellular eukaryotes. This review aims to stimulate discussions on the possibility that the Trypanosomatid flagellum could act as a sensory organ through the MAP kinase pathway, with the objective to encourage investigation of this new hypothesis through a series of proposed experimental approaches.


Activation of the CXCR4 chemokine receptor enhances biological functions associated with B16 melanoma liver metastasis.
The CXCR4 chemokine receptor plays an essential role in the homing of cells to organs expressing its ligand, CXCL12. CXCR4 expressed on tumor cells might regulate their traffic during metastasis. Here, we investigated whether the activation of CXCR4 on B16 murine melanoma cells regulates biological functions associated with metastasis, in vitro and in vivo. Flow cytometry and PCR analysis showed that B16 constitutively expresses high levels of CXCR4 (CXCR4-B16). Biological assays showed that the activation of CXCR4, by its ligand CXCL12, increases the migration, invasion, and proliferation of CXCR4-B16. AMD3100 significantly inhibited the stimulatory migrating effect induced by CXCL12. Treatment of CXCR4-B16 with CXCL12 increases their adhesion to liver sinusoidal endothelial cell (LSEC) monolayers. LSEC, expressing CXCL12, increased the migration of CXCR4-B16. In a liver metastasis model, CXCR4-B16 metastasis was associated with an increased expression of CXCL12 in LSEC territories. CXCR4-B16 cells were located close to LSEC microenvironments expressing CXCL12. Increased liver metastasis was observed after injecting CXCR4-B16 cells previously treated with CXCL12. Our results provide evidence showing that CXCR4 plays an important role in regulating biological functions associated with B16 liver metastasis.


Transcranial magnetic stimulation over the cerebellum and eye movements: state of the art.
Transcranial magnetic stimulation (TMS) transiently induces an electrical field in the tissues beneath the area of application, thereby perturbing local cortical activity if applied over the scalp. It can therefore be used to modulate cerebellar function in healthy humans. Even though the role of the cerebellum in eye movement control and adaptation is well known, few experiments have used eye movements to evaluate the effect of TMS over the cerebellum. Single-pulse TMS over the posterior vermis resulted in impaired accuracy of reflexive saccades, acceleration of smooth pursuit, and coordination of saccades and head movements. TMS over the cerebellar hemisphere decreased pursuit gain. Repetitive TMS (rTMS) over the posterior vermis impaired saccade adaptation in a double-step paradigm. Comparing the effects of TMS on different behavioural paradigms could be useful to test cerebellar control of reflexive and voluntary eye movements, and as a probe of cerebellar plasticity. rTMS appears to be especially interesting since its effects outlast the stimulation period and its behavioural consequences can therefore be measured without interfering with the execution of eye movements or with the experimental procedures.


Evaluation of polygalacturonase activity in Saccharomyces cerevisiae wine strains.
A total of 61 S. cerevisiae strains, 60 of them isolated from wine ecosystems, were evaluated for the presence of the gene encoding endopolygalacturonase (PGU1) and for polygalacturonase (PG) activity. Nine strains lack the gene PGU1 and did not exhibit PG activity on plate assays. Of the 52 strains showing an amplified band corresponding to the size of PGU1 gene, only 36 degraded polygalacturonic acid (PGA) and 17 did not degrade it at any of the pH values used. The coding region of the PGU1 gene (ORF YJR153w) was not present in some PG activity negative strains. The S. cerevisiae UCLMS-39 strain was selected for its specific activity at different pHs, temperatures and oenological parameters. The temperature and pH optima were 50 degrees C and 3.5-5.5 respectively and it was only affected by ethanol. The PGU1 gene was cloned and sequenced. The production of a biologically functional endoPG in S. cerevisiae UCLMS-39 brings us a step closer to improving the qualities of outstanding enological yeasts naturally lacking PG activity.


The effects of lidocaine and procaine on microRNA expression of adipocyte-derived adult stem cells.
The microRNA (miRNA) pathway has emerged as one of the biologic pathways implicated in stem cell regulation. miRNA is a noncoding, single-stranded RNA consisting of 20-25 nucleotides that inhibits the protein production at the step of translation. The molecular effects of lidocaine and procaine on adipose stem cells were investigated by examining RNA expression array. Adipose stem cells were isolated from a prior abdominal liposuction procedure. The human adipose stem cells were cultured and then added to a mixture of 1 ml of culture medium plus 1 ml of 2% lidocaine or 2% procaine for the duration of 30 minutes. The expression levels of miRNAs were estimated by using peptide nucleic acid (PNA)-miRNA array analysis throughout the denaturation and hybridization processes after the isolation of miRNA. The miRNAs detected by microarray that either decreased by half fold or increased by 1.5 fold from the control level were interpreted as significant. According to microarray analysis there were 61 miRNAs in total, and no miRNA had decreased expression levels. The stem cells treatment with lidocaine showed 4 alteration of expression with miR-9a(*) (1.53 fold), miR-29a (1.64 fold), miR-296-5p (1.64 fold) and miR-373 (1.94 fold). The stem cells treated with procaine showed 32 miRNAs that were significantly up-regulated with a range of 1.5 to 2.06 fold. They were stem cell differentiation-related miRNAs, apoptosis and cell cycle-associated miRNAs, immunity-associated miRNAs and hormonal response-related miRNAs. Lidocaine and procaine affect the miRNA expression on adipose stem cells and the effect of procaine is more marked than that of lidocaine.


[Day care, the day care patient and his partner].
The aim of this contribution is to examine what the consequences are of an admission to a psychogeriatric dayhospital for a patient and his partner. In the first place attention has been payed to some complicating factors: on the one side the organization and function of daycare among other institutions, on the other side, at greater length, the direct care to a patient and his partner. Then, an account has been given of a pilotstudy regarding the experience of a patient and his partner in case of an admission to a psychogeriatric dayhospital. By way of literature research, information from family- and patient-meetings and Grid-data, it has been found that it is significant to involve the family in an over-all treatment, in the first place because family can give relevant supplying information; in the second place because an admission can be problematic to such an extent that a partner needs support too. Besides, the carrying-capacity of the family is of crucial importance in case of a daycare-situation. It was further put that more research has to be done into the psychological processes of an admission.


Conserved loop I of U5 small nuclear RNA is dispensable for both catalytic steps of pre-mRNA splicing in HeLa nuclear extracts.
The function of conserved regions of the metazoan U5 snRNA was investigated by reconstituting U5 small nuclear ribonucleoprotein particles (snRNPs) from purified snRNP proteins and HeLa or Xenopus U5 snRNA mutants and testing their ability to restore splicing to U5-depleted nuclear extracts. Substitution of conserved nucleotides comprising internal loop 2 or deletion of internal loop 1 had no significant effect on the ability of reconstituted U5 snRNPs to complement splicing. However, deletion of internal loop 2 abolished U5 activity in splicing and spliceosome formation. Surprisingly, substitution of the invariant loop 1 nucleotides with a GAGA tetraloop had no effect on U5 activity. Furthermore, U5 snRNPs reconstituted from an RNA formed by annealing the 5' and 3' halves of the U5 snRNA, which lacked all loop 1 nucleotides, complemented both steps of splicing. Thus, in contrast to yeast, loop 1 of the human U5 snRNA is dispensable for both steps of splicing in HeLa nuclear extracts. This suggests that its function can be compensated for in vitro by other spliceosomal components: for example, by proteins associated with the U5 snRNP. Consistent with this idea, immunoprecipitation studies indicated that several functionally important U5 proteins associate stably with U5 snRNPs containing a GAGA loop 1 substitution.


Inhibition of the Ca²⁺-dependent K⁺ channel, KCNN4/KCa3.1, improves tissue protection and locomotor recovery after spinal cord injury.
Spinal cord injury (SCI) triggers inflammatory responses that involve neutrophils, macrophages/microglia and astrocytes and molecules that potentially cause secondary tissue damage and functional impairment. Here, we assessed the contribution of the calcium-dependent K⁺ channel KCNN4 (KCa3.1, IK1, SK4) to secondary damage after moderate contusion lesions in the lower thoracic spinal cord of adult mice. Changes in KCNN4 mRNA levels (RT-PCR), KCa3.1 protein expression (Western blots), and cellular expression (immunofluorescence) in the mouse spinal cord were monitored between 1 and 28 d after SCI. KCNN4 mRNA and KCa3.1 protein rapidly increased after SCI; double labeling identified astrocytes as the main cellular source accounting for this upregulation. Locomotor function after SCI, evaluated for 28 d in an open-field test using the Basso Mouse Scale, was improved in a dose-dependent manner by treating mice with a selective inhibitor of KCa3.1 channels, TRAM-34 (triarylmethane-34). Improved locomotor function was accompanied by reduced tissue loss at 28 d and increased neuron and axon sparing. The rescue of tissue by TRAM-34 treatment was preceded by reduced expression of the proinflammatory mediators, tumor necrosis factor-α and interleukin-1β in spinal cord tissue at 12 h after injury, and reduced expression of inducible nitric oxide synthase at 7 d after SCI. In astrocytes in vitro, TRAM-34 inhibited Ca²⁺ signaling in response to metabotropic purinergic receptor stimulation. These results suggest that blocking the KCa3.1 channel could be a potential therapeutic approach for treating secondary damage after spinal cord injury.


Site-specific introduction of an acetyl-lysine mimic into peptides and proteins by cysteine alkylation.
Protein acetylation on Lys residues is recognized as a significant post-translational modification in cells, but it is often difficult to discern the direct structural and functional effects of individual acetylation events. Here we describe a new tool, methylthiocarbonyl-aziridine, to install acetyl-Lys mimics site-specifically into peptides and proteins by alkylation of Cys residues. We demonstrate that the resultant thiocarbamate modification can be recognized by the Brdt bromodomain and site-specific antiacetyl-Lys antibodies, is resistant to histone deacetylase cleavage, and can confer activation of the histone acetyltransferase Rtt109 by simulating autoacetylation. We also use this approach to obtain functional evidence that acetylation of CK2 protein kinase on Lys102 can stimulate its catalytic activity.


Plexogenic arteriopathy in broiler lungs: Evaluation of line, age, and sex influences.
Plexiform lesions form in the terminal pulmonary arterioles of human patients suffering from prolonged pulmonary arterial hypertension. Plexiform lesions also develop in broiler lungs, but lesion incidences are not strongly correlated with sustained pulmonary hypertension as reflected by right to total ventricular weight (RVTV) ratios. The present study was conducted to assess plexiform lesion incidences in broiler lines that have been divergently selected for susceptibility or resistance to pulmonary hypertension. Broilers from susceptible (SUS) and resistant (RES) lines were reared together and only clinically healthy (nonascitic, noncyanotic) individuals were evaluated to minimize potential line differences in cardiopulmonary hemodynamics. The objective was to determine if an innate genetic predisposition for plexogenic arteriopathy would be exposed in SUS broilers when compared with RES broilers in the absence of extreme differences in cardiopulmonary hemodynamics. Broilers up to 12 wk age from the SUS and RES lines had essentially equivalent BW, indices of cardiopulmonary function (left ventricle + septum weight, total ventricle weight, and RVTV ratios), and lung volumes within a sex. Average RVTV ratios for broilers from both lines were indicative of normal pulmonary arterial pressures at all ages sampled. Nevertheless, plexiform lesions were detected in SUS and RES broiler lungs immediately posthatch and thereafter at all ages sampled. Lesion incidences were consistently low and did not differ between the lines within any of the sampling ages. This evidence demonstrates that plexiform lesions develop extremely rapidly in broiler chicks, apparently without the prerequisite for vascular stress caused by severe, prolonged pulmonary arterial hypertension. No innate genetic predisposition for complex vascular lesion development appeared to exist in the SUS line when compared with the RES line.


Carthamus tinctorius flower extract prevents H2O2-induced dysfunction and oxidative damage in osteoblastic MC3T3-E1 cells.
The flowers of Carthamus tinctorius L. (Compositae) have been widely used for enhancing blood circulation and postmenopausal disorder in women. In the present study, the potential protective effects of C. tinctorius flower extract (CFE) against reactive oxygen species (ROS) induced osteoblast dysfunction were investigated using osteoblastic MC3T3-E1 cells. The osteoblast function was assessed by measuring alkaline phosphatase activity, collagen content, calcium deposition, and RANKL production, and the oxidative status was assessed by measuring intracellular lipid peroxidation, and protein oxidation in osteoblastic MC3T3-E1 cells. A significant reduction in the alkaline phosphatase activity, collagen, and calcium deposition and an increase in the production of receptor activator of nuclear factor-kB ligand (RANKL) were observed after 0.3 mM H(2)O(2) addition. The H(2)O(2)-induced alterations were prevented by pre-incubating the osteoblasts with 2-10 microg/ml CFE for 48 h. When the oxidative stress was induced by H(2)O(2), the increased production of protein carbonyl and malondialdehyde was also reduced at the same CFE concentration. These results demonstrate that C. tinctorius flower can act as a biological antioxidant in a cell culture experimental model and protect osteoblasts from oxidative stress-induced toxicity.


Molecular markers for patients with thymic malignancies: not feasible at present?
Thymomas and thymic carcinomas are rare malignancies and devising clinically effective molecular targeted therapies is a major clinical challenge. The aim of the study was to analyze BLC2 and vascular endothelial growth factor receptor (VEGFR) expression and KRAS and EGFR mutational status and to correlate them with the clinical characteristics of patients with thymomas and thymic carcinomas. A total of 62 patients (mean age: 50.4 ± 13.2 years) with thymomas and thymic carcinomas were enrolled. The expression of BLC2 and VEGFR in tumor cells and normal tissues was evaluated by RT-PCR. The mutational status of the KRAS and EGFR genes was investigated by PCR with sequence specific primers. The BLC2 and VEGFR expression levels did not differ significantly between tumor and normal tissues. Moreover, there were no clearly pathogenic mutations in KRAS or EGFR genes in any tumor. None of the molecular markers were significantly related to clinical outcomes. Changes in levels of expression of BLC2 and VEGFR do not appear to be involved in thymic tumorigenesis. Moreover, our data suggest that KRAS and EGFR mutations do not play a major role in the pathogenesis of thymomas and thymic carcinomas.


Gateway RFP-fusion vectors for high throughput functional analysis of genes.
There is an increasing demand for high throughput (HTP) methods for gene analysis on a genome-wide scale. However, the current repertoire of HTP detection methodologies allows only a limited range of cellular phenotypes to be studied. We have constructed two HTP-optimized expression vectors generated from the red fluorescent reporter protein (RFP) gene. These vectors produce RFP-tagged target proteins in a multiple expression system using gateway cloning technology (GCT). The RFP tag was fused with the cloned genes, thereby allowing us localize the expressed proteins in mammalian cells. The effectiveness of the vectors was evaluated using an HTP-screening system. Sixty representative human C2 domains were tagged with RFP and overexpressed in HiB5 neuronal progenitor cells, and we studied in detail two C2 domains that promoted the neuronal differentiation of HiB5 cells. Our results show that the two vectors developed in this study are useful for functional gene analysis using an HTP-screening system on a genome-wide scale.


Lactoferrin protects against acetaminophen-induced liver injury in mice.
Acetaminophen-induced liver injury (AILI) is a significant health problem and represents the most frequent cause of drug-induced liver failure in the United States. The development and implementation of successful therapeutic intervention strategies have been demanding, due to significant limitations associated with the current treatment for AILI. Lactoferrin (Lac), a glycoprotein present in milk, has been demonstrated to possess a multitude of biological functions. Our study demonstrated a profound protective effect of Lac in a murine model of AILI, which was not dependent on its iron-binding ability, inhibition of acetaminophen (APAP) metabolism, or a direct cytoprotective effect on hepatocytes. Instead, Lac treatment significantly attenuated APAP-induced liver sinusoidal endothelial cell dysfunction and ameliorated hepatic microcirculation disorder. This protective effect of Lac appeared to be dependent on hepatic resident macrophages (Kupffer cells [KCs]). Collectively, our data indicate that Lac, through activation of KCs, inhibited APAP-induced liver sinusoidal endothelial cell damage and improved hepatic congestion, thereby protecting against AILI. These findings reveal the significant therapeutic potential of Lac during AILI and other types of liver diseases.